Figure 2. IRES-mediated translation of Nfil3.

(a) Schematic diagram of the bicistronic reporter vector containing the full-length Nfil3 5′-UTR. In the pRF vector, the Nfil3 5′-UTR sequence was inserted between Renilla luciferase and firefly luciferase. Inverse sequences of Nfil3 5′-UTR are also shown. SV40, simian virus 40. (b) MC3T3-E1 cells were transfected with the bicistronic reporter constructs and incubated for 24 hours. Cells were harvested at the indicated times, and cell lysates were subjected to a luciferase assay. Graph shows the ratio of relative luciferase activities, FLUC/RLUC. The ratio of empty vector (pRF) was arbitrarily set to 1.0. Error bars represent mean ± SEM (n = 3), *** P < 0.001.